一种新的用于删除选择标记基因的Cre/lox系统

设计了一种新的诱导型Cre/lox系统,并在转基因烟草(Nicotianatabacum L.)中进行了验证.在诱导剂的作用下,位于同向lox位点之间的选择标记基因(hpt)和重组酶基因(Cre)在烟草愈伤组织中被删除.在该系统中,Cre基因在玉米乙酰苯胺类化合物诱导启动子(In5-2)的控制下表达.对转基因后代的分子检测结果表明,不论是否加入了诱导剂,目的基因(gus)均被整合到烟草基因组中;在诱导剂处理的48株转基因烟草To代中,45株的hpt基因被删除了.该系统只使用一个载体,克服了二次转化系统带来的问题.

A New Cre/lox System for Deletion of Selectable Marker Gene

A new inducible Cre/lox system was constructed in transgenic tobacco (Nicotiana tabacum L.) plants. The inducer-treatment of tobacco callus mediates an excision event in which the selectable marker gene and Cre gene between two lox sites were deleted. A chloroacetanilide-induced promoter (In5-2) was used to control the expression of Cre gene in this system. Molecular analysis of transgenic tobacco plants showed the interested gene, b-glucuronidase (gus), was integrated into the genome whether removing has been successful, and forty-five out of forty-eight T₀ plants were transgenic tobacco without the marker gene, hpt. This system uses a single vector to circumvent the flaw of other dual recombinase vector systems.